The three microelectrode voltage clamp technique will be used to investigate membrane charge movement (gating current) in frog skeletal muscle fibers. In addition, the intracellular free Ca 2 ion concentration change will be measured during step depolarizations using the metallochromic indicator dye arsenazo III. The dye will be injected iontophoretically into fibers in the cutaneous pectoris muscle. A 50 microsm diameter spot of light from a tungsten-halogen lamp will be focused on the point of voltage control. Changes in light transmittance will be measured at three wavelengths using a compound microscope, beam splitters and interference filters. A microscope photometer system with the required sensitivity has been constructed and tested. The objective of the research is to test the hypothesis that membrane charge movement in muscle is a gating current for Ca 2 ion release from the sarcoplasmic reticulum. The hypothesis will be tested by examining whether charge movement and the Ca 2 ion transient behave in parallel during a variety of experimental conditions. For example, the voltage dependence, subthreshold response, threshold behavior and the response to prolonged depolarization will be examined.